Elastic Fiber Staining Research Articles

Biallelic and monoallelic variants in EFEMP1 can cause a severe and distinct subtype of heritable connective tissue disorder.

Variants in EFEMP1, encoding Fibulin-3, were previously reported as a rare cause of heritable connective tissue disorder (HCTD) with recurrent hernias and joint hypermobility. We report three new cases with biallelic or monoallelic EFEMP1 variants and severe hernia phenotypes. Two male siblings of 10 and 13 years old presented with marfanoid habitus, recurrent inguinal and umbilical hernias, generalized joint hypermobility, and scoliosis. Parents and halfsiblings reported joint hypermobility and umbilical hernias. The eldest boy died at age 16 from incarcerated gastrointestinal herniation complicated by gastric and bowel necrosis with perforation. Autopsy revealed widespread intestinal diverticula. Immunohistochemistry of skin and fascia tissue did not reveal any abnormalities, including normal staining of elastic fibers. Both siblings harbored compound heterozygous likely pathogenic EFEMP1 variants (c.1320 + 2T > A, p.? and c.698G > A, p.Gly233Asp). An unrelated 58-year-old male had marfanoid features, high myopia, recurrent diaphragmatic and inguinal hernias, and chronic gastrointestinal dilatation with severe malabsorption. Both his dizygotic twin-brother and mother had recurrent hernias and high myopia. This man died at 59 years of age, and autopsy showed extensive diaphragmatic herniation, bowel diverticula, and pulmonary emphysema. A heterozygous EFEMP1 splice-variant (c.81 + 1G > A, p.?) was identified, causing exon skipping leading to a start-loss. Targeted genome reanalysis nor RNA-sequencing revealed a second variant at the other allele. The reported individuals expand the clinical and pathological phenotypes of EFEMP1-related disease, a distinct entity within the spectrum of HCTD. The severe and recurrent hernias, gastrointestinal dilatation, and diverticulosis result in an increased risk for life-threatening complications, demanding early recognition and close monitoring.

Baseline computed tomography imaging findings could assist in early diagnosis of visceral pleural invasion for newly discovered early subpleural non-small cell lung cancer: T1 or T2.

Preoperative accurate visceral pleural infiltration (VPI) diagnosis for T1-size non-small cell lung cancer (NSCLC) is significant for clinical decision-making. The study aimed to explore the diagnostic efficacy of computed tomography (CT) imaging features and serum biomarkers in diagnosing VPI in newly discovered subpleural NSCLC ≤3 cm. There were 447 patients with NSCLC ≤3 cm retrospectively enrolled and assigned to the VPI group (n=81) and the non-VPI group (n=366) based on elastic fiber staining results. The serum biomarkers and CT imaging features were obtained for each subject. Univariate and multivariate analyses were used to identify the independent predictors for VPI. Area under the receiver operating characteristic (ROC) curve (AUC) was used to evaluate the diagnostic performance of each independent predictor and combined predictors in predicting VPI, with performance compared using the DeLong test. For tumor biomarkers, the VPI group had a significantly higher percentage of cases with abnormal carcino-embryonic antigen (CEA) level, cytokeratin 19 fragment (CYFRA21-1) level, and pro-gastrin-releasing peptide (ProGRP) level than that of the non-VPI group (P<0.001, P=0.003, P=0.004). However, in multivariate analysis, only the lesion-pleura relationship patterns type Ia [odds ratio (OR) =20.689; 95% confidence interval (CI): 5.058-84.622; P<0.001], type Ib (OR =5.155; 95% CI: 1.178-22.552; P=0.03), type II (OR =7.154; 95% CI: 1.733-29.53; P=0.007) with type III as reference, solid lesion density (OR =9.954; 95% CI: 4.976-19.911; P<0.001) with part-solid density as reference were identified as the independent predictors for VPI. In predicting VPI, the combined model (AUC =0.885) significantly outperformed models based on lesion density (AUC =0.833) and lesion-pleura relationship patterns (AUC =0.655) (all P<0.001). The CT predictors for VPI in patients with subpleural NSCLC (≤3 cm) were lesion density and lesion-pleura relationship patterns (pleural attachment and indentation), but not serum tumor biomarkers.

Facteurs pronostiques : degré d’invasion de la sous-séreuse, distance tumeur-séreuse et invasion de la limitante élastique de la sous-séreuse dans l’adénocarcinome colique

The aim was to study the prognostic impact of tumor infiltration of the subserosa in colonic adenocarcinoma, by evaluating the degree of tumor infiltration in the subserosa (DISS), tumor-serosa distance (DTS), and invasion of the elastic boundary of the subserosa (ILE) after elastic fiber staining.Material and methods: All patients operated on for colonic adenocarcinoma classified as pT3 without lymph node or visceral metastasis operated on at CHU-Amiens between 2004 and 2017 were included. All slides were reviewed by 2 pathologists. Bivariate and subgroup analyses were performed according to the presence of a DISS ≤5 mm or >5mm, a DTS ≤1mm or >1mm and the presence or absence of an ILE. These statistical analyses were then correlated with 5-year survival.Results: 101 patients were included in the study. We performed elastic fiber staining on an average of 2 tumor blocks per case, and 39.6% of patients had invasion of the elastic boundary. However, bivariate and subgroup analyses showed no statistically significant association between DISS, DTS or ILE and 5-year survival.Conclusion: None of these three histological criteria proved to have prognostic value in our series, contrary to some results in the literature. However, as these data are subject to a number of confounding factors, we do not recommend that pathologists specify these different criteria in their reports.

Clinicopathological Appearance of Epidermal Growth-Factor-Containing Fibulin-like Extracellular Matrix Protein 1 Deposition in the Lower Gastrointestinal Tract: An Autopsy-Based Study.

This study examined the patterns of epidermal growth-factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1) deposition in the small intestine and colon to evaluate the association between the histopathological severity of EFEMP1 deposition and constipation and determine the colocalization of amyloid transthyretin (ATTR) and EFEMP1 deposits. In 40 older cases (≥80 years of age), EFEMP1 deposition in the small intestine initiated in the submucosal and subserous vessels, subserous interstitium, and serosa (early stage), progressing to the muscularis propria and peri-Auerbach plexus area (intermediate stage), and finally spreading diffusely to other areas, excluding the mucosa and muscularis mucosa (advanced stage). The colon had a similar pattern of progression. During the middle-to-advanced stages, amyloid formation was observed in some vascular and serous deposits. A subgroup of cases was identified in which EFEMP1 deposition was the only presumed cause of constipation. Additionally, we demonstrated the colocalization of ATTR and EFEMP1 deposition. Apple-green birefringence was detected under polarized light only in approximately one-half of the cases in the small intestine and one-third of the cases in the colon. These findings strongly suggest that EFEMP1 deposits are correlated with pathological conditions of the lower gastrointestinal tract. As the histopathological diagnosis using Congo red-stained specimens is challenging, the combined use of elastic fiber staining and EFEMP1 immunohistochemistry is recommended to identify EFEMP1 deposition.

Facial Skin Rejuvenation Using Poly- dl -Lactic Acid Injected With a Laser-Generated Needle-Free Microjet Injector.

A laser-induced needle-free microjet injector was developed for rapid, high-speed drug delivery of microliters into the skin. This study evaluated the clinical rejuvenation effect of repeated dermal injections of the collagen simulator poly- dl -lactic acid (PDLA) using a laser-induced needle-free microjet injector. Five PDLA injection sessions using a laser-induced needle-free microjet injector were conducted in patients concerned about aging skin. Facial uplifting, darkness, redness, roughness, pore size, subjective satisfaction, and side effects were evaluated before each session and 4 weeks after treatment completion. Histological evaluation was also performed with immunohistochemical staining of collagen and elastic fibers. The clinical results of 27 female patients were evaluated. The treatment resulted in a noticeable skin surface uplifting (0.711 ± 0.42 mm) and significant improvements in darkness ( p = .013), redness ( p = .009), and roughness ( p = .036), with no significant difference in the pore size ( p = .770). Patients were reported being satisfied with the overall therapeutic effects, despite mild and tolerable adverse effects. Histological findings revealed growth and thickening of collagen and elastic fibers, with marked increase in collagen I and III levels. Repeated dermal injections of PDLA using a laser-induced microjet injector offer excellent drug delivery, achieving high efficacy in skin rejuvenation, patient satisfaction, and safety.

Dynamics of structural transformations of intervertebral discs in humans in the fetal period

BACKGROUND:The process of formation of the intervertebral disc structure remains poorly understood. Therefore, evaluating the forecasts for the development of the disc’s fibrous component as the foundation of its strength and elastic properties in the postnatal period seems appropriate.&#x0D; AIM:To identify microstructural transformations of the annulus fibrosus and nucleus pulposus in the fetal period and compare them with the ontogenetic orientation of the spinal motion segment.&#x0D; MATERIAL AND METHODS:The study material included 150intervertebral discs obtained during autopsy of 50fetuses. The gestational age of 42fetuses was in the early fetal period, 8fetuses were in the late fetal period. Intervertebral discs СV–СVI, ThV–ThVI, LV–SIwere examined in each fetus. All histological preparations were stained with hematoxylin and eosin, silver impregnation method, PAS reaction, Alcian blue (pH=1.0), Van Gieson and Weigert stains. For intergroup comparisons, the Kruskal–Wallis, Mann–Whitney, and Newman–Keuls tests were used. Differences were considered statistically significant atp≤0.05.&#x0D; RESULTS:Elastic fibers in the late fetal period were found in the annulus fibrosus and the peripheral zone of the nucleus pulposus. The intensity of staining of collagen and elastic fibers was more pronounced in the outer layers of the annulus fibrosus. Analysis of the parameters of the vascular-connective tissue formations showed an increase in their number (p=0.0368) and an increase in the number of vessels in the vascular-connective tissue formations of the intervertebral disc (p=0.0449) in the direction of the lower levels of the spinal motion segments. Differences in these two indicators were obtained between the СV–СVIand LV–SIdiscs, as well as between the СV–СVIand ThV–ThVIdiscs. In relation to all the studied morphometric parameters of the vascular-connective tissue formations in the cervical and thoracic spine, no statistically significant differences were obtained between the early and late fetal periods.&#x0D; CONCLUSION:The development of the intervertebral disc occurs from the peripheral parts to the center; the source for its further development is the fibrous ring.

Optimized elastic fiber staining with sodium dichloroisocyanurate and BRCA1 double staining in diagnosis of lung cancer.

Optimized elastic fiber staining with sodium dichloroisocyanurate and BRCA1 double staining in diagnosis of lung cancer.

Hepatic vein anatomy related to Laennec's capsule for liver resection.

Laennec's capsule is the proper membrane covering the entire surface of the liver parenchyma, including around the Glissonean pedicles and hepatic veins. Laennec's capsule around the hepatic veins has been clinically reported to comprise two layers: the hepatic and cardiac Laennec's capsules. However, where the cardiac Laennec's capsule is derived from and the two capsules' separability are still unclear. This study aimed to investigate the anatomy of the two capsules using cadaveric specimens. Histopathological examinations of autopsy specimens from four cadavers were conducted. Each specimen was sliced in the longitudinal section to include the pericardium, right hepatic vein (RHV), and inferior vena cava (IVC). Additionally, long-axis specimens of the RHV were obtained to determine the separability of the capsules and to examine the extent to which the capsules extended along the RHV. Laennec's capsule was estimated using elastic fiber staining. The cardiac Laennec's capsule is derived from the pericardium (parietal and visceral) and diaphragm and runs parallel with the IVC and hepatic veins associated with the hepatic Laennec's capsule. The two capsules were separable micro- and macroscopically. The hepatic Laennec's capsule was observed from the root to the peripheral side and the cardiac Laennec's capsule from the root to the middle side of the hepatic vein. The existence of the two layers of Laennec's capsule around the hepatic veins was confirmed histologically and they were separable. Identification and decollement of these layers would contribute to establishing safe and feasible anatomic liver resection techniques.

Prediction of Axillary Lymph Node Metastasis in Invasive Breast Cancer by Sound Touch Elastography.

The aims of this study were to investigate the value of sound touch elastography (STE) in predicting axillary lymph node metastasis (ALNM) in patients with invasive breast cancer (IBC) and to explore whether lysyl oxidase (LOX) is correlated with increasing stiffness and promotion of metastasis in IBC. A total of 142 lesions in 142 patients were assessed by STE. The STE values of IBCs in the two groups were compared and the best cutoff values for diagnosing ALNM determined. Immunohistochemistry was used to detect LOX expression. Collagen fiber and elastic fiber content was determined by Masson and Weigert elastic fiber staining. Correlation analyses were performed to identify the associations of the data. The optimal cutoff values of Emax (maximum stiffness value of the tumor) and Smax (maximum stiffness value of the shell) for predicting ALNM of IBC were 94.58 and 148.78 kPa. Immunohistochemistry and Masson and Weigert elastic fiber staining were performed on 67 samples. LOX expression and collagen volume fraction were significantly higher in the ALNM+ group than in the ALNM- group (p=0.04 and 0.03), except for elastic fiber content (p=0.628). Moreover, Emax, Smax and LOX expression were positively correlated with collagen volume fraction (r=0.624, 0.512, and 0.533, respectively). Emax and Smax were found to be predictors for ALNM of IBC. STE could serve as a non-invasive method for assessing lymph node status before surgery. Overexpression of LOX and increased collagen fiber contributed to the increased stiffness in the lesions and metastases of IBC.

Staining method for coronary arteries

Detection of elastic and collagen fibers in the same histological section to determine their relationships and quantification is important for connective tissue investigations. The aim of the study was to assess the possibility of differentiated staining of elastic and collagen fibers with resorcin-fuchsin and fast green FCF in the same histological section and compare the result of this method with the results of the classical methods of staining connective tissue, performed according to standard protocols. Material and methods. We studied adult human hearts, obtained from patients who died from non-cardiac causes. We cut tissue sections from the anterior interventricular artery and the underlying myocardium. After formalin fixation, standard processing and embedding in paraffin of the material, histological sections were made. Histological sections were deparaffinized and immersed resorcin-fuchsin for 15 minutes, then stained with Weigert’s iron hematoxylin for 2 minutes, then for 2 minutes were applied to the preparation surface 0.1% solution of fast green FCF mixed with a saturated solution of picric acid immediately before staining in a ratio of 1:10, the histological sections were dehydrated, cleared and mounted in permanent mounting medium. Results. Elastic fibers were dyed in dark blue, and collagen fibers in different shades of green, which made it possible to differentiate them on the basis of color differences in one histological section. Conclusion. We have developed and tested the method of staining, which provides the possibility of simultaneous detection of elastic and collagen fibers with successive staining with solutions of resorcin-fuchsin and fast green FCF on one histological section.

Splenic Artery Embolization and Splenectomy for Spontaneous Rupture of Splenic Hemangioma and Its Imaging Features.

BackgroundSpontaneous splenic rupture (SSR) is a rare, often life-threatening, acute abdominal injury that requires immediate diagnosis and early treatment. SSR is mainly treated surgically or conservatively. A few cases of interventional embolization for SSRs have been reported.Case PresentationA 30-year-old male patient complaining mainly of left upper abdominal pain underwent emergency abdominal computed tomography (CT) and showed enlargement of the spleen with a massive mixed-density shadow approximately 10.0 × 8.0 × 12.5 cm in size. The boundary was unclear and showed obvious progressive enhancement. Considering the intrasplenic tumor lesions with rupture and hemorrhage, the possibility of vascular tumors was high, with intraperitoneal blood and fluid accumulation. Digital subtraction angiography of the splenic arteriography and embolization of the ruptured splenic artery branches were performed. Postoperative hemoglobin progressively decreased, inflammatory indicators, such as white blood cell counts, procalcitonin (PCT), and C-reactive protein (CRP) were significantly increased, and 2 days after embolization, the patient developed severe hypoxemia, shock, pulmonary edema, and acute respiratory distress syndrome. CT re-examination 9 days after embolization showed reduced lesion absorption. After stabilization of the condition, splenectomy was performed, and postoperative platelet count increase, anticoagulant improvement, and discharge were observed. Postoperative pathological examination revealed extensive hemorrhage and necrosis, vascular tissue with abnormal hyperplasia in the surrounding area, vascular tissue in the bleeding area and outer wall (elastic fiber staining +), and local myofibroblast hyperplasia. Immunohistochemistry showed actin (SM +) and Ki67 (10% +).ConclusionSSR caused by splenic hemangioma is rare, and the choice between surgical treatment or splenic artery embolization remains dependent on the patient's hemodynamic stability and imaging findings.

Insulin Resistance Promotes the Formation of Aortic Dissection by Inducing the Phenotypic Switch of Vascular Smooth Muscle Cells.

BackgroundInsulin resistance (IR) plays a key role in the development of type 2 diabetes mellitus (T2DM) and is one of its most important characteristics. Previous studies have shown that IR and T2DM were independent risk factors for a variety of cardiovascular and cerebrovascular diseases. However, there are few studies on the relationship between IR and aortic dissection (AD). The goal of this research was to find evidence that IR promotes the occurrence of AD.MethodsThrough the statistical analysis, we determined the proportion of glycosylated hemoglobin (HbA1c) abnormalities (HbA1c > 5.7) in people with acute thoracic aortic dissection (ATAD) and compared the difference of messenger RNA (mRNA) and protein expression of GluT1 in the thoracic aorta of normal people and those with ATAD to find evidence that IR is a causative factor in AD. The mouse model of IR and AD and the IR model of human aortic vascular smooth muscle cells (HA-VSMC) were established. Real time-PCR (RT-PCR) and Western blotting were used to study the mRNA and protein expression. Hematoxylin and eosin (H&E), Masson, and elastic fiber staining, and immunofluorescence were used to study the morphological structure.ResultsThe proportion of HbA1c abnormalities in patients with ATAD was 59.37%, and the mRNA and protein expression of GluT1 were significantly lower than that in normal people. Fasting glucose concentration (FGC), serum insulin concentration (SIC), and the homeostasis model assessment of insulin resistance (HOMA-IR) of mice was obviously increased in the high-fat diet group and the protein expressions of Glut1 and GluT4 were reduced, indicating that the mouse IR model was successfully established. The incidence of AD was different between the two groups (IR: 13/14, Ctrl: 6/14), and the protein expression of MMP2, MMP9, and OPN were upregulated and SM22 and α-SMA were downregulated in mice. The expressions of mRNA and protein of GluT1 and SM22 in HA-VSMCs with IR were reduced and OPN was increased.ConclusionCombined results of clinical findings, mouse models, and cell experiments show that IR induced the phenotypic switching of vascular smooth muscle cells (VSMCs) from contractile to synthetic, which contributes to the occurrence of AD. It provides a basis for further research on the specific mechanism of how IR results in AD and a new approach for the prevention and treatment of AD.

Anterior cruciate ligament innervation in primary knee osteoarthritis.

To relate the Anterior Cruciate Ligament (ACL) innervation and histologic degeneration status to the knee osteoarthritis radiologic and functional status. Prospective observational study including 30 consecutive patients affected by primary knee osteoarthritis undergoing Total Knee Arthroplasty (TKA). All patients suffering secondary knee osteoarthritis, an antecedent of an infectious process, malignant process, autoimmune disorder, or previous knee surgery were excluded. We recorded biodemographic, clinical, and radiologic variables of all participants previous to the TKA procedure. ACL tissue was harvested during TKA standard procedure and the obtained sample was fixed in 4% formalin and paraffin-embedded. ACL cross-sections were stained by haematoxylin-eosin and Gallego staining for elastic and collagen fibers, and Sevier-Munger silver staining for nervous tissue. ACL samples histologic degeneration classification reported 15.4% normal, 23.1% slight, 26.9% mild, 11.5% moderate and 23.1% marked. We noted 46.2% large nervous fascicles, 15.4% medium fascicles, 3.8% small fascicles, and no nerve fibers were found in 34.6% ACL samples. No significant correlation was found between the histologic degeneration and the nervous fiber quantification (p>0.05, in all cases). We noted a significant histologic degeneration inverse correlation with the VAS scale (p=0.016), and nervous fiber quantification correlation with Lequesne maximum distance walked punctuation (p=0.043). We also noted greater nervous fiber quantification with minor radiological knee osteoarthritis (Kellgren-Lawrence grade II). ACL degeneration and innervation deficit may play a role in primary knee osteoarthritis onset, but the lack of a defining relationship among the different parameters assessed justifies further research in greater populations.

Salvianolic acid B alleviates diabetic endothelial and mitochondrial dysfunction by down-regulating apoptosis and mitophagy of endothelial cells

ABSTRACT Endothelial dysfunction is a critical mediator in the pathogenesis of vascular complications of diabetes. Herein, this study was conducted to investigate the therapeutic effects of Salvianolic acid B (Sal B) on diabetes-induced endothelial dysfunction and the underlying mechanisms. Diabetic models were established both in db/db mice and high glucose (HG)-induced human umbilical vein endothelial cells (HUVECs). Moreover, HUVECs were exposed to carbonyl cyanide m-chlorophenyl hydrazone (CCCP) to induce endothelial cell damage. Following Sal B treatment, pathological changes of thoracic aorta were investigated by hematoxylin and eosin, alcian blue (AB), elastic fiber, Masson, and reticular fiber staining. BCL2-associated X (BAX), B-cell lymphoma-2 (Bcl-2), Beclin1, Parkin and PTEN Induced Kinase 1 (Pink1) expression was detected by Western blot, immunohistochemistry, and immunofluorescence in thoracic aorta, HG- and CCCP-induced HUVECs. Cell scratch test, MitoTracker Red CMXRos staining and Flou-4 AM staining were separately presented to detect migration, mitochondrial activity and intracellular Ca2+ in HUVECs. Our results showed that Sal B significantly ameliorated hyperlipidemia, hyperglycemia, hyperinsulinemia, and insulin resistance in db/db mice. Furthermore, it significantly alleviated diabetes-induced vascular endothelial dysfunction according to histopathology analysis. In diabetic thoracic aorta, HG- and CCCP-induced HUVECs, Sal B distinctly increased Bcl-2 expression and reduced BAX, Beclin1, Parkin and Pink1 expression, thereby protecting endothelial cells from apoptosis and mitophagy. Moreover, Sal B markedly enhanced migration, mitochondrial activity and intracellular Ca2+ levels both in HG- and CCCP-induced HUVECs. Collectively, Sal B exhibited a potential to improve diabetes-induced endothelial and mitochondrial dysfunction through down-regulating apoptosis and mitophagy of endothelial cells. Abbreviations: DM: diabetes mellitus; T2DM: type 2 diabetes mellitus; Sal B: Salvianolic acid B; HG: high glucose; FBG: fasting blood glucose; TC: total cholesterol; TG: triglycerides; LDL-C: low-density lipoprotein cholesterol; HDL-C: high-density lipoprotein cholesterol; FINS: fasting insulin; HOMA-IR: homeostasis model assessment insulin resistance; QUICKI: quantitative insulin-sensitivity check index; H&E: hematoxylin and eosin; HUVECs: human umbilical vein endothelial cells; IHC: immunohistochemistry; CCCP: carbonyl cyanide m-chlorophenyl hydrazone; FCM: flow cytometry; CCK-8: cell counting kit-8

Compensatory role of endogenous sulfur dioxide in nitric oxide deficiency-induced hypertension

ObjectiveThis study aimed to determine the communicational pattern of gaseous signaling molecules sulfur dioxide (SO2) and nitric oxide (NO) between vascular endothelial cells (VECs) and vascular smooth muscle cells (VSMCs), and elucidate the compensatory role and significance of endogenous SO2 in the development of hypertension due to NO deficiency. Approach and resultsBlood pressure was monitored by the tail-cuff and implantable physiological signal telemetry in L-nitro-arginine methyl ester (l-NAME)-induced hypertensive mice, and structural alterations of mouse aortic vessels were detected by the elastic fiber staining method. l-NAME-treated mice showed decreased plasma NO levels, increased SO2 levels, vascular remodeling, and increased blood pressure, and application of l-aspartate-β-hydroxamate, which inhibits SO2 production, further aggravated vascular structural remodeling and increased blood pressure. Moreover, in a co-culture system of HAECs and HASMCs, NO from HAECs did not influence aspartate aminotransferase (AAT)1 protein expression but decreased AAT1 activity in HASMCs, thereby resulting in the inhibition of endogenous SO2 production. Furthermore, NO promoted S-nitrosylation of AAT1 protein in HASMCs and purified AAT1 protein. Liquid chromatography with tandem mass spectrometry showed that the Cys192 site of AAT1 purified protein was modified by S-nitrosylation. In contrast, dithiothreitol or C192S mutations in HASMCs blocked NO-induced AAT1 S-nitrosylation and restored AAT1 enzyme activity. ConclusionEndothelium-derived NO inhibits AAT activity by nitrosylating AAT1 at the Cys192 site and reduces SO2 production in HASMCs. Our findings suggest that SO2 acts as a compensatory defense system to antagonize vascular structural remodeling and hypertension when the endogenous NO pathway is disturbed.

An analytical study of neocartilage from microtia and otoplasty surgical remnants: A possible application for BMP7 in microtia development and regeneration.

To investigate auricular reconstruction by tissue engineering means, this study compared cartilage regenerated from human chondrocytes obtained from either microtia or normal (conchal) tissues discarded from otoplasties. Isolated cells were expanded in vitro, seeded onto nanopolyglycolic acid (nanoPGA) sheets with or without addition of bone morphogenetic protein-7 (BMP7), and implanted in nude mice for 10 weeks. On specimen harvest, cartilage development was assessed by gross morphology, histology, and RT-qPCR and microarray analyses. Neocartilages from normal and microtia surgical tissues were found equivalent in their dimensions, qualitative degree of proteoglycan and elastic fiber staining, and quantitative gene expression levels of types II and III collagen, elastin, and SOX5. Microarray analysis, applied for the first time for normal and microtia neocartilage comparison, yielded no genes that were statistically significantly different in expression between these two sample groups. These results support use of microtia tissue as a cell source for normal auricular reconstruction. Comparison of normal and microtia cells, each seeded on nanoPGA and supplemented with BMP7 in a slow-release hydrogel, showed statistically significant differences in certain genes identified by microarray analysis. Such differences were also noted in several analyses comparing counterpart seeded cells without BMP7. Summary data suggest a possible application for BMP7 in microtia cartilage regeneration and encourage further studies to elucidate whether such genotypic differences translate to phenotypic characteristics of the human microtic ear. The present work advances understanding relevant to the potential clinical use of microtia surgical remnants as a suitable cell source for tissue engineering of the pinna.

Modified Intravascular Interventional Operation of Balloon-Bound Elastase Induction Method to Establish Rabbit Aneurysm Model Using Cardiovascular Angiography

Objective: In this paper, we establish a simple, efficient and stable rabbit aneurysm model using modified intravascular interventional operation of balloon-bound elastase induction method. Methods: We selected 20 healthy adult New Zealand white rabbits. An aortic aneurysm model of rabbit right common carotid artery was established by modified intravascular interventional balloon-binding elastase induction method using cardiovascular angiography. The cerebral angiography confirmed the model successfully after 4 weeks. Some animals were given paraffin-embedded H-E staining and elastic fiber staining after sacrifice. Angiography was performed for all experiments to build our models and to confirm our findings. Results: Two rabbits died during postoperative feeding, and the remaining 18 rabbits were cultured for 4 weeks and confirmed by angiography to make an aneurysm model. The success rate of aneurysm was 90%, the diameter of the tumor-bearing artery was 3.12±0.20, the short diameter of the tumor was 3.41±0.88, the long diameter of the tumor was 6.30±1.65, and the width of the neck was 3.49±0.80. HE pathology showed that the destruction of the elastic layer of the aneurysm disappeared, similar to the pathological changes of human aneurysms. Conclusions: The rabbit model of aneurysm was established by modified intravascular intervention combined with elastase induction based on angiography. It is similar to human aneurysm in morphology, histology and hemodynamics. The method is simple, the damage is small, the aneurysmal formation rate is high, and the performance is high. Stable and reliable, low production cost, it is an ideal animal model for the development of preclinical neurological interventional devices.

The sympathetic transmitter norepinephrine inhibits VSMC proliferation induced by TGFβ by suppressing theexpression of the TGFβ receptor ALK5 in aorta remodeling.

The sympathetic system is involved in the arterial diseases, but its mechanism remains poorly understood. The present study aimed to explore the impact of the sympathetic neurotransmitter norepinephrine (NE) on transforming growth factor (TGF) β signaling and the role of NE in aortic remodeling. Guanethidine was used to induce a regional chemical sympathetic denervation (CSD) in angiotensin II (AngII) and β-aminopropionitrile (BAPN)-induced aortic aneurysm models. The diameter of the aorta was measured, and elastic fiber staining was performed. TGFβ type I receptor kinase (ALK5) expression in rat aortic NE-treated vascular smooth muscle cells (VSMCs) was detected by reverse transcription-quantitative PCR and western blotting. The effects of NE and ALK5 overexpression on migration, proliferation, apoptosis and TGFβ signaling were also evaluated. Furthermore, adrenergic receptor blockers were used to determine which receptor was involved in the modulation on TGFβ signaling by NE. The results of the present study demonstrated that CSD protected rats from AngII+BAPN-induced aortic remodeling and aneurysm formation. Compared with the control group, NE inhibited VSMC proliferation and migration, but promoted apoptosis by suppressing ALK5 expression, reversing the effects of TGFβ signaling through the suppression of the SMAD-dependent canonical pathway and promotion of the non-canonical pathway. These effects were prevented by ALK5 overexpression. The inhibition of α- or β-adrenergic receptors alleviated the NE-mediated suppression of ALK5 expression. In conclusion, regional CSD protected rats from aortic aneurysm. NE inhibited SMAD2/3-dependent TGFβ signaling by suppressing ALK5 expression, which may serve an important role in VSMC biological functions. Both α- and β-adrenergic receptors were involved in the regulation of ALK5 expression by NE. Abnormal sympathetic innervation of the aorta may be used as a therapeutic target in aortic diseases.

Effect of Long-term Administration of Prostaglandin E1 on Morphologic Changes in Ductus Arteriosus

To improve survival of patients with hypoplastic left heart syndrome, combination therapy with bilateral pulmonary artery banding and prostaglandin E1 (PGE1)-mediated ductal patency was developed as an alternative for high-risk neonates in Japan. However, the effect of long-term PGE1 administration on ductus arteriosus remains unclear. Synchrotron radiation-based X-ray phase-contrast tomography (XPCT) enables clear visualization of soft tissues at an approximate spatial resolution of 12.5 μm. We aimed to investigate morphologic changes in ductus arteriosus after long-term PGE1 infusion using XPCT. Seventeen ductus arteriosus tissue samples from patients with hypoplastic left heart syndrome were obtained during the Norwood procedure. The median duration of lipo-prostaglandin E1 (lipo-PGE1) administration was 48 days (range, 3 to 123). Structural analysis of ductus arteriosus was performed and compared with conventional histologic analysis. The XPCT was successfully applied to quantitative measurements of ductal media. Significant correlation was found between the duration of lipo-PGE1 infusion and mass density of ductal media (R= 0.723, P= .001). The duration of lipo-PGE1 administration was positively correlated with elastic fiber staining (R= 0.799, P < .001) and negatively correlated with smooth muscle formation (R=-0.83, P < .001). No significant increase in intimal cushion formation was found after long-term lipo-PGE1 administration. Expression of ductus arteriosus dominant PGE2-receptor EP4 almost disappeared in specimens when lipo-PGE1 was administered over 3 days. Disorganized elastogenesis and little intimal cushion formation after long-term lipo-PGE1 administration suggest that ductus arteriosus remodeled to the elastic artery phenotype. Because EP4 was downregulated and ductus arteriosus exhibited elastic characteristics, the dosage of lipo-PGE1 might be decreased after a definite administration period.

丘疹性弹性纤维离解22例临床病理分析

Objective To analyze clinical and pathological features of papular elastorrhexis. Methods Clinical data were collected from 22 patients with confirmed papular elastorrhexis in Hospital for Skin Diseases, Chinese Academy of Medical Sciences and Pekin Union Medical College from September 2006 to May 2018. Clinical manifestations, pathological findings and follow-up results were retrospectively analyzed. Results The average age of onset of the 22 patients was 5.7 years (range: 1 - 10 years) , and the male to female ratio was 4.5∶1. The average duration from the occurrence of disease to the confirmation of diagnosis was 1.5 years, and no definite etiology was found. The patients had no itching or pain sensation. Skin lesions were soft, slightly elevated, well-circumscribed, round, oval or polygonal-shaped, white papules with diameters of 1 - 10 mm, and wrinkles appeared on the surface of the papule when the papule was pushed towards its center. Among the 22 patients, 16 (73%) presented with scattered lesions, 13 (59%) had less than 5 papules, and lesions were located in the trunk in 21 (95%) . Histopathological examination of skin lesions in 8 patients showed no obvious increase of collagen fibers in the superficial and middle dermis, which were normally arranged with slightly widened spaces between them. Elastic fiber staining showed that elastic fibers disappeared or were dissociated focally in the superficial and middle dermis. After confirmed diagnosis, the 22 patients received no treatment. In 18 patients, skin lesions did not continue to expand after onset, and no new skin lesions occurred. Skin lesions were slightly enlarged, but remained steady thereafter in 4 patients. Sixteen patients achieved partial remission. Conclusions Papular elastorrhexis is a rare skin disorder of elastic fibers that occurs predominantly during childhood and adolescence, and its diagnosis relies on clinical manifestations combined with histopathological findings. No special treatment is needed and the prognosis is good. Key words: Elastic tissue; Skin manifestations; Papular elastorrhexis